Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae Transcription Units

ACS Synthetic Biology
Neta AgmonJef D Boeke

Abstract

We have adapted the Golden Gate DNA assembly method to the assembly of transcription units (TUs) for the yeast Saccharomyces cerevisiae, in a method we call yeast Golden Gate (yGG). yGG allows for the easy assembly of TUs consisting of promoters (PRO), coding sequences (CDS), and terminators (TER). Carefully designed overhangs exposed by digestion with a type IIS restriction enzyme enable virtually seamless assembly of TUs that, in principle, contain all of the information necessary to express a gene of interest in yeast. We also describe a versatile set of yGG acceptor vectors to be used for TU assembly. These vectors can be used for low or high copy expression of assembled TUs or integration into carefully selected innocuous genomic loci. yGG provides synthetic biologists and yeast geneticists with an efficient new means by which to engineer S. cerevisiae.

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Citations

May 10, 2015·Nucleic Acids Research·Leslie A MitchellJef D Boeke
Oct 22, 2016·ACS Synthetic Biology·Simon John MoorePaul S Freemont
Dec 23, 2016·Nucleic Acids Research·Lauren B A WoodruffChristopher A Voigt
Nov 24, 2017·Frontiers in Microbiology·Shuobo Shi, Huimin Zhao
Nov 23, 2018·Science China. Life Sciences·Zhouqing LuoJunbiao Dai
May 24, 2018·Nature Communications·Lena HochreinBernd Mueller-Roeber
May 24, 2018·Nature Communications·Yi WuJef D Boeke
Nov 21, 2019·Nucleic Acids Research·Neta AgmonJef D Boeke
Mar 12, 2020·Current Protocols in Molecular Biology·Sylvestre Marillonnet, Ramona Grützner
Jun 23, 2020·Annual Review of Biochemistry·Weimin ZhangJef D Boeke
Feb 9, 2017·Proceedings of the National Academy of Sciences of the United States of America·Neta AgmonJef D Boeke
Jul 14, 2019·Scientific Reports·David ChiassonMartin Parniske
Feb 22, 2017·Microbial Biotechnology·Ewelina CelińskaJean-Marc Nicaud
Dec 1, 2018·Nature Communications·Sonja BillerbeckVirginia W Cornish
Jan 29, 2017·Nucleic Acids Research·Lena HochreinBernd Mueller-Roeber
Dec 12, 2020·Frontiers in Microbiology·Claire L Gordy, Carlos C Goller
Sep 14, 2019·Biotechnology Advances·Antoine VassauxValérie Leclère
Dec 5, 2020·Nature Communications·Meng ZhaoJef D Boeke
Aug 11, 2021·Microbial Cell Factories·Weiwang DouWenjun Guan
Feb 20, 2019·ACS Synthetic Biology·Duo LiuYing-Jin Yuan
Aug 28, 2021·ACS Synthetic Biology·Weibo QiaoYansheng Zhang

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